1. Down-Regulation of Telomerase Activity and Activation of Caspase-3 Are Responsible for Tanshinone I-Induced Apoptosis in Monocyte Leukemia Cells in Vitro
- Author
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Rui-Fang Fan, Jia-Jun Liu, Yong Zhang, Ren-Wei Huang, Xiao-Dan Liu, Zhi-Gang Fang, Heqing Huang, Ruo-Zhi Xiao, Dong-Jun Lin, Peiqing Liu, Wei-Bing Guan, and Hong-Zhi Yang
- Subjects
Telomerase ,Survivin ,Tanshinone I (Tan-I) ,telomerase ,survivin ,leukemia ,Down-Regulation ,Apoptosis ,Biology ,Catalysis ,Article ,Inhibitor of Apoptosis Proteins ,lcsh:Chemistry ,Inorganic Chemistry ,Western blot ,Cell Line, Tumor ,medicine ,Humans ,Telomerase reverse transcriptase ,MTT assay ,Viability assay ,Physical and Theoretical Chemistry ,lcsh:QH301-705.5 ,Molecular Biology ,Spectroscopy ,Cell Proliferation ,medicine.diagnostic_test ,Caspase 3 ,Gene Expression Regulation, Leukemic ,Organic Chemistry ,General Medicine ,U937 Cells ,Molecular biology ,Antineoplastic Agents, Phytogenic ,Computer Science Applications ,Blot ,Enzyme Activation ,lcsh:Biology (General) ,lcsh:QD1-999 ,Abietanes ,Cancer research ,Monocytic leukemia - Abstract
Tanshinone I (Tan-I) is a diterpene quinone extracted from the traditional herbal medicine Salvia miltiorrhiza Bunge. Recently, Tan-I has been reported to have anti-tumor effects. In this study, we investigated the growth inhibition and apoptosis inducing effects of Tan-I on three kinds of monocytic leukemia cells (U937, THP-1 and SHI 1). Cell viability was measured by MTT assay. Cell apoptosis was assessed by flow cytometry (FCM) and AnnexinV/PI staining. Reverse transcriptase polymerase chain reaction (RT-PCR) and PCR-enzyme-linked immunosorbent assay (ELISA) were used to detect human telomerase reverse transcriptase (hTERT) expression and telomerase activity before and after apoptosis. The activity of caspase-3 was determined by Caspase colorimetric assay kit and Western blot analysis. Expression of the anti-apoptotic gene Survivin was assayed by Western blot and Real-time RT-PCR using the ABI PRISM 7500 Sequence Detection System. The results revealed that Tan-I could inhibit the growth of these three kinds of leukemia cells and cause apoptosis in a time- and dose-dependent manner. After treatment by Tan-I for 48 h, Western blotting showed cleavage of the caspase-3 zymogen protein with the appearance of its 17-kD subunit, and a 89-kD cleavage product of poly (ADP-ribose) polymerase (PARP), a known substrate of caspase-3, was also found clearly. The expression of hTERT mRNA as well as activity of telomerase were decreased concurrently in a dose-dependent manner. Moreover, Real-time RT-PCR and Western blot revealed a significant down-regulation of Survivin. We therefore conclude that the induction of apoptosis by Tan-I in monocytic leukemia U937 THP-1 and SHI 1 cells is highly correlated with activation of caspase-3 and decreasing of hTERT mRNA expression and telomerase activity as well as down-regulation of Survivin expression. To our knowledge, this is the first report about the effects of Tan-I on monocytic leukemia cells.
- Published
- 2010